PCRmax Leishmania tropica Instruction Manual

Leishmania tropica mspC gene
for GP63

Leishmania tropica

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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Leishmania tropica is a single-celled trypanosome parasite belonging to the genus
Leishmania, which causes cutaneous leishmaniasis in humans. It is commonly found along
the Mediterranean Coast, Middle East, Central Africa and certain parts of Asia. Its lifecycle
encompasses two morphological states; the promastigote which is a motile flagellate that
is approximately 10-20 µm in length and the amastigote which is spheroid to ovoid in
shape and approximately 2.5- 5.0 µm in length. It has a double stranded, linear DNA
genome comprising 36 chromosomes ranging in size from 0.35 to -3 Mb.

There are two hosts in the Leishmania’s protozoan life-cycle. The intermediate host is the
female sand-fly. In the mid-gut of the sand-fly, the parasite is a promastigote that
reproduces asexually and moves to the pharynx. From here it is expelled into the skin of a
vertebrate host when bitten. The parasite is initially phagocytosed by macrophages before
spreading to blood and tissue cells of the host. Once within the host cell, the flagellum is
lost and the parasite, in its amastigote form, replicates by binary fission. The host cell
eventually bursts and the parasites are released to infect other cells throughout the body.

Cutaneous leishmaniasis can be identified by the large number of skin ulcers it produces
within few weeks of being bitten by female sand-flies. These ulcers are noticeably present
on the exposed part of the body such as the face, arms and legs. On healing, these ulcers
leave disabling scars. Patients suffering from this disease tend to develop secondary
infections that could potentially be fatal.

Introduction to Leishmania tropica

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MIN

MAX

Our kit for human Leishmania tropica has been designed for the specific and exclusive in
vitro quantification of this species and does not detect other species within this subgenus.
The primers and probe sequences in this kit have 100% homology with a broad range of
clinically relevant reference sequences based on a comprehensive bioinformatics
analysis.

The PCR Max qPCR Kit for Leishmania tropica (L.tropica) genomes is designed for the in
vitro quantification of L.tropica genomes. The kit is designed to have the broadest
detection profile possible whilst remaining specific to the L.tropica genome.

The primers and probe sequences in this kit have 100% homology with a broad range of L.
tropica sequences based on a comprehensive bioinformatics analysis.

If you require further information, or have a specific question about the detection profile of
this kit then please send an e.mail to [e-mail] and our bioinformatics team will
answer your question.

Specificity

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Kit Contents

• L.tropica specific primer/probe mix (150 reactions BROWN)

FAM labelled

• L.tropica positive control template (for Standard curve RED)

• Internal extraction control primer/probe mix (150 reactions BROWN)

VIC labelled as standard

• Internal extraction control DNA (150 reactions BLUE)

• Endogenous control primer/probe mix (150 reactions BROWN)

FAM labelled

• RNAse/DNAse free water (WHITE)

for resuspension of primer/probe mixes and internal extraction control DNA

• Template preparation buffer (YELLOW)

for resuspension of positive control template and standard curve preparation

Reagents and equipment to be supplied by the user

Real-Time PCR Instrument

DNA extraction kit

This kit designed to work well with all processes that yield high quality DNA with minimal
PCR inhibitors.

Lyophilised 2x qPCR Mastermix

This kit is designed to be compatible with all commercially available Mastermixes that run
with standard cycling conditions.

Pipettors and Tips

Vortex and centrifuge

Thin walled 1.5 ml PCR reaction tubes

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